Hyperbaric Criogenesis Chambers

ABSTRACT

The Hyperbaric Cryogenesis Chambers are equipment for medical usage, extracorporeal, and capable of promoting the proliferation and preservation of cells in a way mainly microbiologic, “not biologic”. They are compartments with containers that host tissues or cells in solution with nutrients, that bear pressures higher and lower to sea level They have a cold system inside that lowers the environmental temperature and maintains it permanently. They are for maintaining viability of cells or tissues and to induce their organized proliferation with oxygen or other gases at pressures higher or lower that the exterior of the chamber.

Description of the patent that for a period of 20 years, is applied by the Medical Doctor Gastón Jerónimo VERRI LIMA, holder of I.C. 1.504.703-1, Montevideo, Uruguay, for an invention of his property named: Hyperbaric Criogenesis Chamber, the creation whose patent is applied for consists of a compartment with the capacity to be pressurized to pressures superior to the equivalent of an Atmosphere of pressure (760 mmHg) and depressurized in an intermittent way, with whatever mixture of gases as pure oxygen, compressed air, Nitrox, Heltiox, Trimix, and/or anaesthetic gases. Inside it there is a system which contains in a liquid or semi-liquid element: cells, complete or incomplete organic tissues, complete organs or fractions of them, and/or human bodies or dead animals. They have inside or outside an hermetic system in order to lower the environment temperature. It may have an illumination system, optic and microscopic visualization, optic or chamber fibroscopy; environment temperature sensors. It has pressure manometers and over-pressure safety valves. The sizes are variable.

There are no known solutions regarding the combination of induction in the cellular proliferation in an extra-corporeal manner and in an organized way, in one or a few cellular types maintaining cellular integrity, provoking the descent of the temperature in compartments which hold them with physiologic fluids under intermittent pressure with pure oxygen. There are no known solutions, either, which combine both events in order to induce the no-proliferation and/or lysis of tumoral cells by direct action of free radicals of oxygen under conditions of raised pressure and of cold.

The crio-preservation of tumoral cells and the induction in the no-proliferation or in the lysis of tumoral cells is a technique used since more or less two decades by hemotherapists and in hemato-oncology; but with a limitation of time, because this preservation is not permanent. Hyiperbaric oxygenation has been used for more than three decades in order to accelerate the wound cicatrisation of patients, by introducing them in hyperbaric chambers and inhale pure oxygen in an intermittent way under an environment pressure higher than sea level. (one atmosphere or 760 mmHG.) for a period that may reach an hour and twenty minutes, and so managing to accelerate the speed of cellular reproduction in the affected area in an organized way. So, said proliferation of cells of determined types is not realized in an extra-corporeal manner.

The advantages in the use of Hyperbaric Criogenesis Chambers is establishing a combination of both techniques in order to induce reproduction of cells of a cellular type that although being mixed with tumoral cells, we get to process them, viability is maintained for a longer period of time, and at the same time it fosters the lysis or the no-proliferation of tumoral cells at a higher speed than these techniques allow separately; but furthermore in an extracorporeal manner. This combination of crio-preservation and hyperbaric oxygen-therapy with the processing in Hyperbaric Criogenesis Chambers, accelerates cellular reproduction of various types determining the possibility of an implant when the number of these cells is already insufficient; as an example in leukemia in its terminal phase and the hepato-pathology with cirrhosis in its terminal stage, among other pathologies. Besides, this invention, with the adequate technique, allows the preservation of human corpses or dead animals for longer periods.

To build Hyperbaric Criogenesis Chambers is necessary to build chambers apt to be pressurized to pressures up to 10 Kg per square centimeter with certified steel or aluminum plate (pursuant to ISO norms on Hyperbaric Chambers types A, B or C). that in its inside or outside has a refrigerating system, characterized by an hermetic steel or aluminum tubing able to bear pressure and permit the installation of another tubing preferably of copper in its inside. The tubing preferably of copper is the one which allows the circulation of refrigerating gas in its inside, isolated from the steel or aluminum by a compacted cotton net. The copper tubing is connected to a motor for the circulation of gas under electric propulsion. The Hyperbaric Criogenesis Chamber can hold receptacles preferably made of glass, named containers, with the capacity to store physiologic fluids and cells, tissues, organs and human bodies or dead animals. So its size is variable.

The industrial exploitation is realized in hospitals with the installation in places with Hyperbaric Criogenesis Chambers in order to process in an extracorporeal manner cells or tissues for the treatment of diseases as leukemias in its final stage or hepato-pathologies with cirrhosis in its terminal stage, among other diseases through obtaining samples of viable cells or tissues in order to realize one or more implants to the patient (that will include, if it is necessary, the mother cells).

In a way not strictly related to hospitals, in everything which deals with the preservation of human bodies or dead animals for larger periods of time.

The Hyperbaric Criogenesis Chambers may be transportable or fixed.

DESCRIPTION OF DRAWING OF HYPERBARIC CRIOGENESIS CHAMBERS

A: Manometers to register the temperature of the internal environment of the Hyperbaric Criogenesis Chamber.

B: Thermometer to register the temperature of the internal environment of the Hyperbaric Criogenesis Chamber. C: Observation System with glass resistant to 10 kg/cm² of pressure or fibroscopy for internal observation with microscopy or chamber connected to a monitor. D: Shutting standard cranks of the door of the Hyperbaric Criogenesis Chamber. E: Cold generator with closed circuit and Inner-chamber System isolated with no static material. F: Supply system of gases at higher pressure interchangeable with pure oxygen or of exclusive supply of pure oxygen under pressure. G: Decompression valves.

H: Exhaust System or Decompression.

I: Compression System with its corresponding valves. J: Hyperbaric Criogenesis Chamber with closed circuit of the refrigerating system and circulating system of gases under pressure. K: Container of physiologic fluids, cells, tissues, organs and bodies, with or without its essential nutrients, with or without micrometric gradation at its bottom, preferably made of glass. L: Overpressure safety valve.

Procedure

The Hyperbaric Criogenesis Chambers allow storage and processing of tissues or organs for the induction of the proliferation of the cellular type in an organized way, that is to say no tumoral. It also permits the induction in the no-proliferation or the lysis of eventual tumoral cells (it will be defined as disorganized) of those partial or complete samples of tissues or organs in accordance with its processing. To achieve this, there must be kept isolated the samples of cells with nucleus of whatever type (including mother cells, epithelial, pancreatic, neurons, fibroblasts, hepatic cells, muscular cells, ovocytes, spermatozoids, etc.) in receptacles (named containers), as partial or total tissue, or as organ. Temperature is lowered to values less than 37 degrees Celsius, that may surpass −70 degrees Celsius with the same environment pressure to that of sea level (that is to say without modifying the pressure). The isolated cells with nucleus, or in the form of tissues or partial or complete organs, are in receptacles (containers of various sizes according to necessities) with the particularity that remain from the beginning of the process, in a solution with physiologic fluids such as serum, plasma, plasmatic substitutes; in association with essential nutrients (preparations of parentheral nutrition or similars, with or without vitamins and/or minerals), under physiological conditions of PH, and being preserved by the descent in temperature to variable values depending on the cellular type that may surpass −70 degrees Celsius. Afterwards, depending upon the cellular type to be processed, comes the programmed rise in temperature inside the Hyperbaric Criogenesis Chamber and the environment pressurization reaching values higher than an atmosphere of pressure that could reach pressures up to 7 atmospheres. The compression or pressurization is realized with pure oxygen; or also with other gases like compressed air, or mixtures such as anesthetic gases, nitric oxide, Trimix, Heliox, Nitrox, etc., to be substituted by pure oxygen one hundred percent. The time of pressurization can vary and maybe more than an hour, with a sequence during the day also variable that may be once or as many times as necessary in order to obtain the cellular sample according to requirement. Once obtained the time of pressurization pre-established to process the sample, we proceed to decompression and lowering of the temperature inside the Hyperbaric Criogenesis Chamber so as to reach the initial values.

The Hyperbaric Criogenesis Chambers allow with this procedure the induction in the no-proliferation or to the lysis of disorganized cells (tumorals) of whatever sample, by the direct action of cold and the direct action of free radicals of oxygen obtained in conditions of saturation at pressures higher than an atmosphere with said gas (viability disturbance and proliferation of tumoral or disorganized cells by exposition to cold and free radicals).

In the procedure, the Hyperbaric Criogenesis Chambers may be operated manually or by programmed control with computers.

These Hyperbaric Criogenesis Chambers may have or not, an optic or microscopic visualization system, or by way of chambers of the samples through optic fibroscopy.

State of the Art

It is of public knowledge the existence of Hyperbaric Chambers, they have been used for more than one hundred years to help patients in decompression or diving accidents, skin wounds, infections, burns, fractures, compartment syndromes, refractory mycosis, or intoxications, among other diseases. The patient is helped by introducing him in the Hyperbaric Chamber and when it is pressurized inside, and reaches values higher than an atmosphere, pure oxygen is administered for the patient to breathe, so resulting in microbiologic effects. The pressurization is realized with compressed air in the so-called multi-seater chambers, or pure oxygen in single-seaters. In any way pure oxygen is breathed under higher pressure than it is found at sea level utilizing the closed circuit systems in the multi-seater chambers. The Hyperbaric Chambers were classified in three categories as follow: 1) Type A, for human beings and multi-seater. 2) Type B, for human beings and single-seater. 3) Type C, for experimentation and animals.

It is also of public knowledge the handling of samples of tissues or cells in cold conditions so as to preserve the cells, as it has been the case in animal insemination for more than three decades.

It is a novelty the creation of Hyperbaric Criogenesis Chambers taking advantage of the incorporation of both techniques, for the microbiologic processing activity. Processing that in its procedure leads to preservation and induction of organized proliferation of isolated cell samples or as tissues or organs in forms partial or complete, under sequence variations of temperature and pressure, with pure oxygen and nutrients in dissolution in a medium of physiologic fluids which hosts said cells. Also, with the Hyperbaric Criogenesis Chambers we get the induction in the no-proliferation of disorganized or tumoral cells or the lysis of them by the associated action of fire radicals and cold. 

1) The Hyperbaric Criogenesis Chamber. 2) The procedure of usage of the Hyperbaric Criogenesis Chambers. 3) The microbiologic product obtained by the use of the Hyperbaric Criogenesis Chambers. 4) The preservation for unlimited time of the microbiologic product or the biologic sample with the use of the Hyperbaric Criogenesis Chambers. 5) The promotion in the induction of lysis or the inhibition in the proliferation of eventual disorganized or tumoral cells of the sample that is processed with the Hyperbaric Criogenesis Chambers. 6) The potentiality to realize direct and continuous visualization of the microbiologic event induced in the Hyperbaric Criogenesis Chambers. 7) The Hyperbaric Criogenesis Chambers in their capacity as transportable or not. 